BRENDA - Enzyme Database

Molecular cloning, expression, characterization and mutation of Plasmodium falciparum guanylate kinase

Kandeel, M.; Nakanishi, M.; Ando, T.; El-Shazly, K.; Yosef, T.; Ueno, Y.; Kitade, Y.; Mol. Biochem. Parasitol. 159, 130-133 (2008)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.4.8
DNA and amino acid sequence determination and analysis, sequence comparison
Plasmodium falciparum
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.7.4.8
D74A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
2.7.4.8
E101D
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
2.7.4.8
N103D
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
2.7.4.8
Y76F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.4.8
AMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
CMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
dAMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
dCMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
dGMP
competitive versus GMP and mixed-type versus ATP
Plasmodium falciparum
2.7.4.8
EDTA
85% inhibition at 3 mM
Plasmodium falciparum
2.7.4.8
additional information
no inhibition by TMP and by 2-mercaptoethanol up to 5 mM
Plasmodium falciparum
2.7.4.8
UMP
10-15% inhibition at 5 mM
Plasmodium falciparum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.4.8
additional information
-
additional information
kinetics of wild-type and mutant enzymes
Plasmodium falciparum
2.7.4.8
0.0222
-
GMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
0.0592
-
GMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
0.0616
-
GMP
mutant E101D
Plasmodium falciparum
2.7.4.8
0.0731
-
dGMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
0.0746
-
dGMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
0.1816
-
dGMP
mutant D74A
Plasmodium falciparum
2.7.4.8
0.1894
-
dGMP
mutant E101D
Plasmodium falciparum
2.7.4.8
0.1933
-
GMP
mutant D74A
Plasmodium falciparum
2.7.4.8
0.2265
-
GMP
mutant N103D
Plasmodium falciparum
2.7.4.8
0.2364
-
dGMP
mutant N103D
Plasmodium falciparum
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.4.8
ATP + GMP
Plasmodium falciparum
-
ADP + GDP
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.4.8
Plasmodium falciparum
-
-
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.4.8
30
-
with substrates ATP and dGMP
Plasmodium falciparum
2.7.4.8
750
-
with substrate ATP and GMP
Plasmodium falciparum
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.4.8
ATP + AMP
very low activity with AMP
700079
Plasmodium falciparum
ADP + ADP
-
-
-
?
2.7.4.8
ATP + dGMP
low activity, dGMP acts also as inhibitor
700079
Plasmodium falciparum
ADP + dGDP
-
-
-
?
2.7.4.8
ATP + GMP
-
700079
Plasmodium falciparum
ADP + GDP
-
-
-
?
2.7.4.8
additional information
no activity with CMP, dTMP, dAMP, dCMP, and UMP
700079
Plasmodium falciparum
?
-
-
-
-
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.4.8
2
8
dGMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
26.5
-
dGMP
mutant E101D
Plasmodium falciparum
2.7.4.8
26.8
-
dGMP
mutant N103D
Plasmodium falciparum
2.7.4.8
33.1
-
dGMP
mutant D74A
Plasmodium falciparum
2.7.4.8
43
-
dGMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
848
-
GMP
mutant E101D
Plasmodium falciparum
2.7.4.8
946
-
GMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
1032
-
GMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
1080
-
GMP
mutant N103D
Plasmodium falciparum
2.7.4.8
1121
-
GMP
mutant D74A
Plasmodium falciparum
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.4.8
7.2
7.8
-
Plasmodium falciparum
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.7.4.8
6
9
-
Plasmodium falciparum
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.7.4.8
0.148
-
dGMP
versus GMP
Plasmodium falciparum
2.7.4.8
0.4
-
dGMP
versus ATP
Plasmodium falciparum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.4.8
DNA and amino acid sequence determination and analysis, sequence comparison
Plasmodium falciparum
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.4.8
D74A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
2.7.4.8
E101D
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
2.7.4.8
N103D
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
2.7.4.8
Y76F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Plasmodium falciparum
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.4.8
AMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
CMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
dAMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
dCMP
10-15% inhibition at 5 mM
Plasmodium falciparum
2.7.4.8
dGMP
competitive versus GMP and mixed-type versus ATP
Plasmodium falciparum
2.7.4.8
EDTA
85% inhibition at 3 mM
Plasmodium falciparum
2.7.4.8
additional information
no inhibition by TMP and by 2-mercaptoethanol up to 5 mM
Plasmodium falciparum
2.7.4.8
UMP
10-15% inhibition at 5 mM
Plasmodium falciparum
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.7.4.8
0.148
-
dGMP
versus GMP
Plasmodium falciparum
2.7.4.8
0.4
-
dGMP
versus ATP
Plasmodium falciparum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.4.8
additional information
-
additional information
kinetics of wild-type and mutant enzymes
Plasmodium falciparum
2.7.4.8
0.0222
-
GMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
0.0592
-
GMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
0.0616
-
GMP
mutant E101D
Plasmodium falciparum
2.7.4.8
0.0731
-
dGMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
0.0746
-
dGMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
0.1816
-
dGMP
mutant D74A
Plasmodium falciparum
2.7.4.8
0.1894
-
dGMP
mutant E101D
Plasmodium falciparum
2.7.4.8
0.1933
-
GMP
mutant D74A
Plasmodium falciparum
2.7.4.8
0.2265
-
GMP
mutant N103D
Plasmodium falciparum
2.7.4.8
0.2364
-
dGMP
mutant N103D
Plasmodium falciparum
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.4.8
ATP + GMP
Plasmodium falciparum
-
ADP + GDP
-
-
?
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.4.8
30
-
with substrates ATP and dGMP
Plasmodium falciparum
2.7.4.8
750
-
with substrate ATP and GMP
Plasmodium falciparum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.4.8
ATP + AMP
very low activity with AMP
700079
Plasmodium falciparum
ADP + ADP
-
-
-
?
2.7.4.8
ATP + dGMP
low activity, dGMP acts also as inhibitor
700079
Plasmodium falciparum
ADP + dGDP
-
-
-
?
2.7.4.8
ATP + GMP
-
700079
Plasmodium falciparum
ADP + GDP
-
-
-
?
2.7.4.8
additional information
no activity with CMP, dTMP, dAMP, dCMP, and UMP
700079
Plasmodium falciparum
?
-
-
-
-
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.4.8
2
8
dGMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
26.5
-
dGMP
mutant E101D
Plasmodium falciparum
2.7.4.8
26.8
-
dGMP
mutant N103D
Plasmodium falciparum
2.7.4.8
33.1
-
dGMP
mutant D74A
Plasmodium falciparum
2.7.4.8
43
-
dGMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
848
-
GMP
mutant E101D
Plasmodium falciparum
2.7.4.8
946
-
GMP
wild-type enzyme
Plasmodium falciparum
2.7.4.8
1032
-
GMP
mutant Y76F
Plasmodium falciparum
2.7.4.8
1080
-
GMP
mutant N103D
Plasmodium falciparum
2.7.4.8
1121
-
GMP
mutant D74A
Plasmodium falciparum
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.4.8
7.2
7.8
-
Plasmodium falciparum
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.7.4.8
6
9
-
Plasmodium falciparum